3d culture system for culturing cells in synovial fluid, cells cultured in synovial fluid and their use

Abstract

Disclosed herein are materials and methods for culturing cells derived from joint cavities, said culturing being performed in the presence of synovial fluid. Also disclosed herein are cells cultured by the methods described herein, and their use for therapeutic purposes and for identifying therapeutic agents.

What is claimed is: 
     
         1 . A method of culturing mammalian cells that reside in a joint cavity in vivo, comprising:
 a) suspending one or more isolated cells in a three-dimensional matrix; and   b) incubating the suspended cells in a culture medium comprising at least about 20% synovial fluid under conditions suitable for chondrocyte division.   
     
     
         2 . The method of  claim 1 , wherein the mammalian cells are selected from the group consisting of: chondrocytes, synoviocytes, meniscus cells and temporomandibular disc cells. 
     
     
         3 . The method of  claim 2 , wherein the cells are chondrocytes. 
     
     
         4 . The method of  claim 3 , wherein the chondrocytes are obtained commercially or derived directly from a mammalian subject. 
     
     
         5 . The method of  claim 3 , wherein the mammalian chondrocytes are human chondrocytes. 
     
     
         6 . The method of  claim 1 , wherein the culture medium comprises at least about 20% synovial fluid, 30% synovial fluid, 40% synovial fluid, 50% synovial fluid, 60% synovial fluid, 70% synovial fluid, 80% synovial fluid, 90% synovial fluid or 100% synovial fluid. 
     
     
         7 . The method of  claim 1 , further comprising one or more incubations in a culture medium comprising at least about 20% synovial fluid. 
     
     
         8 . The method of  claim 7 , wherein the culture medium is changed at least about every two or three days. 
     
     
         9 . The method of  claim 1 , wherein the three dimensional matrix comprises alginate. 
     
     
         10 . The method of  claim 9 , wherein the three dimensional makes forms a spherical bead about 2 mm in diameter. 
     
     
         11 . The method of  claim 1 , wherein the culture medium is supplemented with CaCl 2 . 
     
     
         12 . The method of  claim 1 , further comprising treating the suspended cells with sodium citrate. 
     
     
         13 . The method of  claim 1 , wherein the one or more suspended cells have been genetically modified. 
     
     
         14 . A method of treating an injury, disease or disorder characterized by chondrocyte depletion comprising administering to a subject afflicted with an injury, disease or disorder characterized by chondrocyte depletion, an effective amount of cultured chondrocytes, wherein the chondrocytes are cultured in at least about 20% synovial fluid and wherein the chondrocytes are suspended in a three-dimensional matrix during culturing. 
     
     
         15 . The method of  claim 14 , wherein the synovial fluid used to culture the chondrocytes is derived from the subject afflicted with the disease or disorder. 
     
     
         16 . The method of  claim 14 , wherein the culture medium comprises at least about 20% synovial fluid, 30% synovial fluid, 40% synovial fluid, 50% synovial fluid, 60% synovial fluid, 70% synovial fluid, 80% synovial fluid, 90% synovial fluid or 100% synovial fluid. 
     
     
         17 . The method of  claim 14 , wherein the three dimensional matrix comprises alginate. 
     
     
         18 . The method of  claim 17 , wherein the three dimensional makes forms a spherical bead about 2 mm in diameter. 
     
     
         19 . The method of  claim 14 , wherein the method is used to treat osteoarthritis. 
     
     
         20 . The method of  claim 14 , wherein the cultured chondrocytes are removed from the three dimensional matrix prior to administering them to the subject. 
     
     
         21 . The method of  claim 14 , wherein the three-dimensional matrix comprises alginate. 
     
     
         22 . The method of  claim 21 , wherein the cultured chondrocytes are removed from the three dimensional matrix prior to administering them to the subject, said removal comprising washing the cells suspended in the matrix with sodium citrate. 
     
     
         23 . The method of  claim 14 , wherein the cultured chondrocytes administered to the subject have been genetically modified. 
     
     
         24 . A mammalian chondrocyte cultured by a method comprising:
 a) suspending one or more chondrocytes in a three-dimensional matrix; and   b) incubating the suspended chondrocytes in a culture medium comprising at least about 20% synovial fluid under conditions suitable for chondrocyte division.   
     
     
         25 . The chondrocyte of  claim 24 , wherein the one or more suspended chondrocytes have been genetically modified. 
     
     
         26 . The chondrocyte of  claim 24 , wherein the three-dimensional matrix comprises alginate. 
     
     
         27 . The chondrocyte of  claim 26 , wherein the chondrocyte is removed from the three-dimensional matrix by washing in sodium citrate. 
     
     
         28 . A method of screening compounds for treating a disease or condition characterized by a depletion of cells in a joint cavity or loss of function of cells in a joint cavity, comprising:
 a) suspending one or more cells in a three-dimensional matrix, wherein the one or more cells are isolated from a subject with a disease or condition characterized by a depletion of cells in a joint cavity or loss of function of cells in a joint cavity;   b) incubating the suspended cells in a culture medium comprising at least about 20% synovial fluid under conditions suitable for chondrocyte division in the presence and/or absence of a test compound; and   c) determining a gene expression profile for one or more genes indicative of a disease state or condition, wherein a gene expression profile of the cells cultured in the presence of the test compound that is more similar to the gene expression profile of a healthy individual than to the gene expression profile of a subject afflicted with the disease or condition indicates the test compound is effective in treating the disease or condition.   
     
     
         29 . The method of  claim 28 , wherein the one or more isolated cells are chondrocytes. 
     
     
         30 . The method of  claim 28 , wherein the disease or condition is osteoarthritis.